雞蛋IgY抗體的生產與純化研究

Abstract

IgY是雞蛋蛋黃中的主要抗體，相較於以哺乳類動物生產IgG多株抗體，以雞蛋生產IgY多株抗體不但方式更為簡便、成本較低廉、產量也較為豐富，且程序也較符合人道要求。此外，由於鳥類與哺乳類動物的親緣關係較遠，IgY在醫學應用上能減少偽陽性反應的發生，因此IgY多株抗體具有基礎研究、一般檢驗、醫學診斷與治療等多方面用途的應用潛力。 在本研究中，我們一方面以小鼠的IgM免疫球蛋白為模式抗原，對雞隻進行免疫注射，以探討生產專一IgY抗體之適當條件；另一方面也發展從液態蛋黃及冷凍蛋黃中粗萃取IgY的方法；此外，我們也利用基因工程的方式，生產抗原的重組蛋白質，並將其製備成吸附管柱，以純化獲得純度、效價及專一性皆優良的IgY抗體樣品。 本研究已成功建立以雞隻生產小鼠免疫球蛋白之IgY抗體的模式，可做為以雞隻生產其他相關抗原之IgY多株抗體的參考；我們也已開發出快速、經濟、有效的IgY萃取與純化方法，除了顯著提升萃取蛋黃IgY的效率與便利性之外，更能增進IgY抗體的應用範圍。IgY is the major antibody in the egg yolk of chicken. There are so many advantages of IgY, the yield in the yolk is abundant, the cost of maintaining hens is low, the using of non-invasive methods to isolate IgY from yolks is more humane, and the false positive result is reduced due to large phylogenetic distance between mammals and birds, so that IgY is a useful tool for basic research, inspection, medical diagnosis and treatment. In this study, we have injected mouse IgM into hens, in order to produce antigen-specific IgY and investigate the appropriate production conditions. On the other hand, we have developed two methods for extracting IgY from liquid form egg yolk and frozen egg yolk respectively. Furthermore, we produced a recombinant antigen protein by genetic engineering, then manufactured an antigen-adsorption column to purify IgY which is high purity, titer and specificity. We have successfully established a production mode of chicken IgY against mouse immunoglobulins, which could be used as a useful reference for the production of IgY antibodies directed to other relevant antigents. We have also developed rapid, economic and effective IgY extraction and purification protocols. These protocols could not only significantly improve the efficiency and the convenience of IgY extraction from egg yolk, but also extend the applicability of IgYantibodies.